Sensitivity of serological and polymerase chain reaction methods for detection of viruses in Allium spp

• Published in: IOP conference series. Earth and environmental science 2020-03, Vol.468 (1), p.12023
• Main Authors: Putri, C A, Hidayat, S H
• Format: Article
• Language: English
• Subjects: Allium; Amplification; Antibodies; Deoxyribonucleic acid; Dilution; DNA; Enzyme-linked immunosorbent assay; Plant extracts; Plant virus diseases; Plant viruses; Polymerase chain reaction; Primers; Sensitivity analysis; Serology; Viruses
• ISSN: 1755-1307; 1755-1315
• DOI: 10.1088/1755-1315/468/1/012023

Infection of Onion yellow dwarf virus (OYDV), Shallot yellow stripe virus (SYSV), and Garlic common latent virus (GCLV) has been reported on Allium spp. in Indonesia. Serology methods using enzyme-linked immunosorbent assay (ELISA), and dot immunobinding assay (DIBA) is a common method to detect viruses, while polymerase chain reaction (PCR) is popular as a molecular method to detect plant viruses nowadays. This research was conducted to assess the sensitivity of 3 detection methods, i.e. ELISA, DIBA and PCR for detecting viruses in Allium spp. Sensitivity level of each method was evaluated by diluting plant extract and antibody for ELISA and DIBA, and cDNA template for PCR. Result of this research showed that DIBA was able to detect OYDV, GCLV, and SYSV with plant extract dilution in the range of 10−1 to 10−2, and antibody dilution from 1:300 to 1:1000. ELISA seems to be more sensitive than DIBA because it was able to detect the same virus with plant extract dilution in the range of 10−1, 10−3, and 10−5, and antibody dilution from 1:300 to 1:1000. RT-PCR method was the most sensitive method compared to ELISA and DIBA because it has the highest accuracy, specification and sensitivity level. Specific DNA fragment was amplified using universal primers for Potyvirus and Carlavirus up to 10−3 and 10−4 dilution factor of cDNA respectively. Moreover, amplification using specific primers of OYDV, SYSV, and GCLV is more sensitive than using universal primers, because up to 10−5 dilution factor of cDNA was able to amplify virus target.